Effects of stromal cell-derived factor-1 on proliferation,migration,and odontoblastic differentiation of human dental pulp stem cells

目的：对比基质细胞衍生因子1（stromal cell-derived factor-1，SDF-1）和粒细胞集落刺激因子（granulocyte colony-stimulating factor，G-CSF）对人牙髓干细胞（dental pulp stem cell，DPSC）的体外增殖、迁移和成牙本质能力的影响。方法：分别在培养基中加入100μg／L SDF-1或100μg／L G-CSF，采用细胞计数试剂盒（cell counting kit-8， CCK-8）和集落形成实验（colony-forming unit，CFU）检测 SDF-1和 G-CSF 对 DPSC 增殖的影响；采用划痕实验和Transwell迁移实验检测两者对DPSC迁移能力的影响；对DPSC进行成牙本质诱导，通过碱性磷酸酶（alkaline phos-phatase，ALP）染色、测定ALP活性、茜素红染色和real-time RT-PCR检测成牙本质相关基因的表达，以检测两者对DPSC体外成牙本质能力的影响。结果：SDF-1和G-CSF能够轻度提高DPSC的增殖及集落形成能力，但差异无统计学意义。加入SDF-1或G-CSF的实验组划痕汇合速率明显高于对照组（P＜0．01），但两种因子间差异无统计学意义。Transwell迁移实验中，对照组每视野的迁移细胞数量为（5．0±1．4）个，SDF-1组每视野的迁移细胞数量为（24．3±6．8）个，G-CSF组每视野的迁移细胞数量为（11．8±3．3）个，各组间差异有统计学意义（P＜0．05）。经成牙本质诱导后，实验组细胞ALP染色加深，ALP活性上升，矿化结节形成数量增加，成牙本质相关基因的表达均显著高于对照组。结论：SDF-1对DPSC的增殖能力影响不显著，但能明显提高DPSC的迁移能力和成牙本质分化能力，效果优于G-CSF。Objective:To compare the effects of stromal cell-derived factor-1 (SDF-1 )and granulocyte colony-stimulating factor (G-CSF)on proliferation,migration,and odontoblastic differentiation of human dental pulp stem cell (DPSC)in vitro.Methods:DPSCs were cultured in vitro and treated with either 1 00 μg/L SDF-1 or 1 00 μg/L G-CSF.Cell counting kit-8 (CCK-8 )and colony-forming unit (CFU ) were used to detect the effect of SDF-1 and G-CSF on the proliferation ability of DPSC.Cell migration of DPSC was determined by wound healing assay and Transwell migration assay.The effects of SDF-1 and G-CSF on odontoblastic differentiation of DPSC were evaluated by alkaline phosphatase (ALP)staining, ALP activity and alizarin red S staining.The expression of odontoblastic-related genes such as dentin ma-trix protein 1 (DMP-1 )and dentin sialophosphoprotein (DSPP)were quantified by real-time RT-PCR. Results:SDF-1 and G-CSF promoted the proliferation of DPSC slightly,but the difference was not statis-tically significant.Wound healing assay showed that SDF-1 and G-CSF promoted cell migration of DPSC significantly (P<0.01 ),but there was no significant difference between the two factors.In Transwell migration assay,the number of migrated cells of the control group was 5 .0 ±1 .4 per sight,while the SDF-1 group was 24.3 ±6.8 per sight and the G-CSF group was 1 1 .8 ±3.3 per sight,suggesting that cell migration of DPSC was improved significantly after being treated with SDF-1 or G-CSF,and SDF-1 was more effective than G-CSF (P<0.05 ).Significantly greater odontoblastic differentiation potential was found in SDF-1 group and G-CSF group based on the ALP staining.Higher ALP activity,more mineralization nodule formation and higher expressions of DMP-1 and DSPP were also found after SDF-1 or G-CSF treatment.Conclusion:SDF-1 had no significant effect on the proliferation of DPSC,but could significantly promote cell migration and odontoblastic differentiation of DPSC.Its effect on DPSC was bet-ter than G-CSF.国家自然科学基金; 北京大学临床医院合作专项(2013-4-01)资助Supported by the National Natural Science Founda-tion of China; the Cooperation Projects of Clinical Hospital of Peking UniversityPubMed中文核心期刊要目总览(PKU)中国科技核心期刊(ISTIC)中国科学引文数据库(CSCD)123-294